Ishihara H, Engel J D, Yamamoto M
Department of Biochemistry, Tohoku University School of Medicine, Sendai.
J Biochem. 1995 Mar;117(3):499-508. doi: 10.1093/oxfordjournals.jbchem.a124736.
To elucidate the mechanisms underlying the tissue-restricted expression of GATA factor transcription, we have isolated and analyzed the genomic chicken GATA-3 (cGATA-3) locus. Structural analysis of the clones showed that the cGATA-3 gene consists of six exons which span more than 19 kb. Two trans-activating domains and two Zn finger domains of cGATA-3 were found to be encoded separately by exons 2/3 and 4/5, respectively, indicating that each functional domain of GATA-3 is encoded by a discrete exon. We have determined 1.7 kb of upstream promoter sequence and found a number of sequence motifs which match those of known transcription factor binding sites. Activities of presumptive regulatory regions of this gene were assessed by transfecting chimeric constructs into a chicken T cell line MSB-1. The results showed three features of cGATA-3 gene regulation. The basal promoter activity of the cGATA-3 gene is determined by sequences lying between -104 and -29 bp of the promoter region. The upstream region containing the GATA and CACCC elements in close proximity (-1280 to -1152) appeared to act as a negative transcriptional regulator, whereas the region -1151 to -850 acts as a positive regulator. Thus, the expression of cGATA-3 gene is under complex regulation and the mode of regulation of cGATA-3 gene expression is suggested to be different from that of GATA-1 genes.
为阐明GATA因子转录的组织限制性表达背后的机制,我们分离并分析了鸡GATA-3(cGATA-3)基因座的基因组。对克隆的结构分析表明,cGATA-3基因由六个外显子组成,跨度超过19 kb。发现cGATA-3的两个反式激活结构域和两个锌指结构域分别由外显子2/3和4/5单独编码,这表明GATA-3的每个功能结构域由一个离散的外显子编码。我们确定了1.7 kb的上游启动子序列,并发现了许多与已知转录因子结合位点相匹配的序列基序。通过将嵌合构建体转染到鸡T细胞系MSB-1中来评估该基因假定调控区域的活性。结果显示了cGATA-3基因调控的三个特征。cGATA-3基因的基础启动子活性由启动子区域-104至-29 bp之间的序列决定。紧邻(-1280至-1152)包含GATA和CACCC元件的上游区域似乎作为负转录调节因子起作用,而-1151至-850区域作为正调节因子起作用。因此,cGATA-3基因的表达受到复杂的调控,并且cGATA-3基因表达的调控模式被认为与GATA-1基因不同。